A high salt diet inhibits obesity and delays puberty in the female rat.

BACKGROUND/OBJECTIVES: Processed foods are considered major contributors to the worldwide obesity epidemic. In addition to high sugar and fat contents, processed foods contain large amounts of salt. Owing to the correlations with rising adiposity, salt has recently been proposed to be obesogenic. This study investigated three hypotheses: (i) high salt contributes to weight gain and adiposity in juvenile female rats, (ii) puberty onset would be altered because salt is known to affect neuronal systems involved in activating the reproductive system, and (iii) enhanced adiposity will act synergistically with salt to drive early puberty onset. DESIGN: Female weanling rats (post-natal day 21, n=105) were fed a low fat/low salt diet, low fat/high salt diet, high fat/low salt diet or a high salt/high fat diet for 24 days. Metabolic measures, including weight gain, food intake, fecal output, activity and temperature were recorded in subsets of animals. RESULTS: Body weight, retroperitoneal and perirenal fat pad weight, and adipocyte size were all lower in animals fed high fat/high salt compared with animals fed high fat alone. Leptin levels were reduced in high fat/high salt fed animals compared with high fat/low salt-fed animals. Daily calorie intake was higher initially but declined with adjusted food intake and was not different among groups after 5 days. Osmolality and corticosterone were not different among groups. Fecal analysis showed excess fat excretion and a decreased digestive efficiency in animals fed high fat/low salt but not in animals fed high fat/high salt. Although respiratory exchange ratio was reduced by high dietary fat or salt, aerobic-resting metabolic rate was not affected by the diet. High salt delayed puberty onset, regardless of dietary fat content. CONCLUSIONS: Salt delays puberty and prevents the obesogenic effect of a high fat diet. The reduced weight gain evident in high salt-fed animals is not due to differences in food intake or digestive efficiency.

Insulin elicits ingestion in decerebrate rats.

Insulin administered to rats reliably elicits ingestion of food. To determine whether the neural mechanisms sufficient to control insulin-elicited ingestion are located in or caudal to the forebrain, decerebrate rats were treated with insulin and ingestive responses were measured. Insulin treatment produced hypoglycemia that was comparable, in magnitude and duration, in control and decerebrate rats. Decerebrate and control rats ingested significantly more sucrose solution while hypoglycemic than while normoglycemic. In contrast, insulin did not augment the water consumption of either group. These data indicate that neural systems caudal to the forebrain are sufficient to control ingestive consummatory behavior through the integration of metabolic signals generated by insulin treatment and taste afferent input from the oropharynx.

Trafficking of tachykinin neurokinin 3 receptor to nuclei of neurons in the paraventricular nucleus of the hypothalamus following osmotic challenge.

Tachykinin neurokinin 3 receptor (NK3R) is a G-protein (GTP binding protein) -coupled receptor that is heavily expressed by magnocellular neurons of the paraventricular nucleus of the hypothalamus (PVN). Osmotic challenge is reported to activate NK3R expressed by magnocellular neurons and cause the NK3R to be internalized to the cytoplasm and perhaps the cell nucleus. In this study we show using immuno-electron microscopy that isolated nuclei from neurons in the PVN of osmotic challenged animals (rats) show a robust labeling for the NK3R. NK3R immunoreactivity was detected by Western blot in isolated nuclei of PVN neurons following the 2 M NaCl injection. No nuclear NK3R immunoreactivity was detected in control animals. NK3R antibody specificity was confirmed by small interfering (SI) RNA technology. This study establishes that the NK3R is trafficked to the nucleus of PVN neurons following a peripheral osmotic challenge.

Applications of taste reactivity to the study of the neural-hormonal controls of ingestive behavior.

Taste plays a central role in guiding ingestive behavior and the encoding of taste is affected by manipulations that influence ingestive behavior. In this article, the use of the taste reactivity test to provide a behavioral assessment of how changes in the oral reinforcing properties of a taste may initiate or sustain ingestive behaviors in several contexts are discussed. The affects of the animal's sex, sodium deficiency, exogenous bombesin administration, and the role of central gustatory lesions in mediating taste reactivity responses are discussed. Findings indicate that an enhancement of ingestive taste reactivity responses correlate with an increased preference and intake of taste stimuli for some, but not all situations. Such situations include the bombesin-like peptides that reduce sucrose and sodium chloride intake without influencing taste reactivity responses. Conversely, female rats, compared to males, show an elevated intake and preference for a range of NaCl concentrations and a greater number of ingestive taste reactivity responses to some, but not all of the preferred concentrations. Such mismatches of taste reactivity and intake measures shift attention to the contribution of nongustatory factors (trigeminal, visceral) in the control of intake.

Distribution of Fos-like immunoreactivity within the rat brain following intraventricular injection of the selective NK(3) receptor agonist senktide.

Neurokinin B (NKB) is one member of an evolutionarily conserved family of neuropeptides, the tachykinins. Preferential binding of NKB to endogenous NK(3) receptors affects a variety of biological and physiological processes, including endocrine secretions, sensory transmission, and fluid and electrolyte homeostasis. In light of its widespread biological actions, immunohistochemical detection of the c-Fos protein product was used to study the distribution of neuronal activation in the rat brain caused by intraventricular (icv) injections of the selective NK(3) receptor agonist (succinyl-[Asp(6), N-Me-Phe(8)] substance P [6-11]), senktide. Quantitative analysis revealed that treatment with isotonic saline or 200 ng senktide resulted in the differential expression of Fos-like immunoreactivity (FLI) throughout the brain. Senktide induced the highest number of FLI neurons in the lateral septum, bed nucleus of the stria terminalis, amygdala, paraventricular nucleus of the hypothalamus, median preoptic nucleus, organum vasculosum of the lamina terminalis, supraoptic nucleus, periaqueductal gray, and medial nucleus of the solitary tract compared to isotonic saline controls. Additional regions that contained elevated FLI following icv injection of senktide, relative to saline injection, included the cerebral cortex, lateral hypothalamic nucleus, suprachiasmatic nucleus, ventral tegmental area, substantia nigra, inferior colliculus, locus coeruleus, zona incerta, and arcuate nucleus. Our data indicate that activation of NK(3) receptors induces the expression of FLI within circumscribed regions of the rat brain. This pattern of neuronal activation overlaps with nuclei known to regulate homeostatic processes, such as endocrine secretion, cardiovascular function, salt intake, and nociception.

Intraventricular injection of tachykinin NK3 receptor agonists suppresses the ingestion of NaCl-associated tastes.

The present experiments evaluated whether a salty taste was required for injections of a neurokinin-3 (NK3) receptor agonist (senktide) to suppress intake or whether senktide would reduce the intake of tastes that are predictive of NaCl. During training, different groups of rats were given access to 1% almond + water, 1% almond + 0.3 M NaCl, or 1% almond + 0.1 M sucrose. On the test day, rats were administered intraventricular injections of either saline or 200 ng senktide and then given access to 1% almond + water. Senktide had no effect on the intake of the water-associated or sucrose-associated almond. In contrast, senktide significantly reduced the intake of NaCl-associated almond. Senktide had no effect on almond intake by water-deprived rats. These results show that activation of NK3 receptors reduces the intake of NaCl and of a neutral taste that is predictive of sodium but not of calories.

Intraoral intake and taste reactivity responses elicited by sucrose and sodium chloride in chronic decerebrate rats.

The oral stimulating effects of sucrose and sodium chloride (NaCl) were assessed in chronic decerebrate and pair-fed intact control rats by measuring both oral motor taste-reactivity responses and intraoral intake volume. Taste-reactivity responses were videotaped during the first minute of the intraoral taste infusion. The infusion continued until the taste solution was rejected from the mouth, and the intake volume was computed accordingly. The number of ingestive taste-reactivity responses and the volume of intraoral intake consumed by pair-fed control and decerebrate rats increased with increasing sucrose concentration. Sucrose intake increased as concentration increased to 0.1 M, then plateaued between 0.3, 1.3, and 2.0 M sucrose for both groups. For control rats, intraoral NaCl elicited an inverted U-shaped function for both taste-reactivity responses and intake. Taste-reactivity responses of chronic decerebrate rats varied with NaCl concentration. In contrast to control rats, intake of NaCl did not differ from that of water for decerebrate rats. These data indicate that caudal brain-stem mechanisms are sufficient to control sucrose intake but are not adequate for the concentration dependent intake of NaCl. Second, these data also indicate that it is possible for taste-elicited oral motor responses to be dissociated from intake. The different roles of taste and postingestive factors in sucrose and NaCl intake are discussed.

Effects of concentration presentation order and intraoral delivery on sucrose intake.

In this article, intraoral intake of an ascending concentration series of sucrose was found to plateau between concentrations of 0.3 M and 2.0 M, and thus failed to show the typical inverted U-shaped intake function found in standard intake tests. Two experiments were conducted to explain this result. In Experiment 1, intraoral and standard 30-min, 1-bottle intake of ascending sucrose concentrations (0.03, 0.1, 0.3, 1.0, 1.3, and 2.0 M) were compared. Sucrose intake was similar for both delivery methods. In a second experiment we examined the effect of the order of sucrose concentration presentation on the 1-bottle 30-min intake of nondeprived intact rats. An ascending concentration order of the solutions produced a significantly greater intake of concentrated sucrose solutions than did a random order. This result strongly suggests that the standard decline in sucrose intake at higher concentrations is determined not only by postoral factors but also by experiential factors (i.e., order of presentation).

Bombesin suppresses need-free and need-induced salt intake in rats.

The researchers performed experiments to evaluate whether the effects of bombesin are selective for the satiation of ingestive behaviors related to energy balance or if ingestive behaviors associated with sodium balance are also suppressed by bombesin. Injections of 4 and 8 micrograms/kg bombesin reliably reduced need-free and sodium deficiency-induced NaCl intake in male rats. The effects of bombesin on the sodium-deficiency-induced change in taste reactivity was assessed. Injections of 4 micrograms/kg and 8 micrograms/kg bombesin had no effect on the sodium deficiency-induced shift in taste reactivity. These data indicate that bombesin suppresses NaCl intake and that bombesin does not appear to interact with gustatory sensibility in exerting its behavior-controlling action.

Inhibition of ingestive behavior following fourth ventricle bombesin injection in chronic decerebrate rats.

To test the hypothesis that the effects of 4th ventricle bombesin (BN) injection on feeding require interaction with forebrain neural systems, intraoral sucrose (0.1 M) was measured in tube-fed control and tube-fed supracollicular decerebrate rats after 4th ventricle injections of 1, 5, 10, 20, and 50 ng BN. Fourth ventricle injections of all doses of BN reliably suppressed sucrose intake in both control and chronic decerebrate rats. These results indicate that caudal brain stem afferent signals produced by 4th ventricle BN injections are integrated by the local neural circuitry of the caudal brain stem, independent of the forebrain systems, to modulate ingestive behavior.

Sodium homeostasis in chronic decerebrate rats.

Physiological and behavioral concomitants of sodium need were studied in supracollicularly transected and pair-fed intact rats. Chronic decerebrate rats, like intact rats, reduced their urine sodium output when placed on a sodium-deficient diet. Similarly, 24 hr after sodium loading, decerebrate and intact rats excreted comparable levels of the excess sodium. In the 2 hr immediately following the loading, decerebrate rats excreted less sodium. In contrast, behavioral aspects of sodium homeostasis were completely absent in chronic decerebrate rats. In separate experiments, intraoral intake, and taste-reactivity responses, elicited by intraoral infusions of NaCl were measured during sodium-replete and sodium-deficient conditions. In response to oral infusions of NaCl, intact rats consumed significantly more and produced greater numbers of ingestive taste-reactivity responses when they were sodium deficient than when they were sodium replete. The same sodium-depletion treatments in chronic decerebrate rats, however, altered neither the intraoral intake of NaCl nor the frequency of NaCl-elicited ingestive taste-reactivity responses. These results suggest that the behavioral compensatory responses that follow changes in the internal sodium state are dependent upon forebrain mechanisms.

Chronic voluntary nicotine drinking enhances nicotine palatability in rats.

The response of rats to nicotine solutions was examined with the brief-exposure, taste reactivity test and a two-bottle, 24-hr preference test. In Experiment 1, naive nondeprived rats were administered intraoral infusions (0.8 ml infused during 1 min) of distilled water and 1 microgram/ml, 5 micrograms/ml, 10 micrograms/ml, 25 micrograms/ml, 50 micrograms/ml, and 100 micrograms/ml nicotine. The oral motor, taste reactivity (TR) responses of the rats were recorded during the infusion. Nicotine solutions up to a concentration of 50 micrograms/ml elicited a number of ingestive TR responses similar to that by water. Ingestive responses significantly decreased and aversive TR responses significantly increased in response to 100 micrograms/ml nicotine. On the basis of these results, two-bottle preferences for water versus 1 microgram/ml, 5 micrograms/ml, and 0 microgram/ml (water control group) nicotine were measured in three groups of naive rats. Rats initially showed an equal preference for 0 microgram/ml and 1 microgram/ml nicotine. After 16 days of exposure, however, rats developed a significant preference for 1 microgram/ml nicotine. The preference ratio for 5 micrograms/ml nicotine significantly increased during the experiment, but the preference ratio remained significantly less than that for 1 microgram/ml and 0 microgram/ml nicotine solutions. Last, TR responses elicited by intraoral infusions of 1 microgram/ml and 5 micrograms/ml nicotine were then measured in these rats having had the two-bottle experience. Rats showing a two-bottle preference for the 1 microgram/ml nicotine solution displayed significantly more ingestive TR responses to 1 microgram/ml and 5 micrograms/ml nicotine than did the control rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Central gustatory lesions: II. Effects on sodium appetite, taste aversion learning, and feeding behaviors.

Intake and taste reactivity tests were used to determine the effects of bilateral lesions of the gustatory portions of the nucleus of the solitary tract (NST), the parabrachial nucleus (PBN), and the ventral posteromedial nucleus of the thalamus (VPMpc) on several complex ingestive behaviors. In the 1st experiment, lesions of the PBN and the NST blocked, and VPMpc lesions impaired, the behavioral expression of salt appetite. In the 2nd experiment, alanine was paired with injections of LiCl. Control rats as well as rats with NST and VPMpc lesions acquired the taste aversion, but rats with PBN lesions did not. In the 3rd experiment, all animals increased their food intake after injections of 2 U/kg insulin and 250 mg/kg 2-deoxy-D-glucose, and their food intake was suppressed after nutritive stomach loads.

Central gustatory lesions: I. Preference and taste reactivity tests.

Bilateral electrophysiologically guided lesions were placed in the nucleus of the solitary tract (NST), the parabrachial nucleus (PBN), and the ventral posteromedial thalamic nucleus (VPMpc) of rats, and 15-min intake and taste reactivity (TR) responses elicited by 3 concentrations each of sucrose, NaCl, HCl, and quinine (Q) HCl were subsequently measured. Compared with controls, NST lesions had no significant effects on intake, and rats with PBN lesions consumed significantly more QHCl, sucrose, NaCl, and HCl. Thalamic lesions decreased sucrose intake. Analysis of TR responses showed that the QHCl threshold for aversive responses increased after VPMpc, PBN, and NST lesions. Rats with NST or PBN lesions were unresponsive to increasing sucrose concentration. TR responses elicited by NaCl and HCl were similar across the groups.

Comparison of the interoceptive sensory consequences of CCK, LiCl, and satiety in rats.

In three experiments we assessed the degree to which ad lib feeding, injection of cholecystokinin (CCK), and injection of lithium chloride (LiCl) produce states with similar sensory consequences. In each experiment, two groups of rats were trained to use cues arising from food deprivation and satiation as discriminative signals for shock. One group was shocked when deprived but not when nondeprived. The other group received the reversed discrimination. Testing began when incidence of freezing was greater under the shocked deprivation than under the nonshocked deprivation condition. In Experiment 1, the rats were tested under 24-hr food deprivation after injections of CCK, LiCl, and saline (in counterbalanced order). We reasoned that if either CCK or LiCl induce satiety-like states, they should promote patterns of responding different from those produced by saline but similar to those produced by ad lib feeding. The effects of CCK on freezing did not differ from those of saline, whereas both CCK and LiCl had effects that were different from ad lib feeding. This pattern of results was also obtained when deprivation level during training and testing was reduced to 8 hr (Experiment 1A) and also when rats received small amounts of food in conjunction with CCK (Experiment 2). The intubation of a high-calorie stomach load (Experiment 1A) produced a response profile like that observed after free feeding. Freezing after LiCl treatment differed from that observed after free feeding and from that found after injection of CCK. The results indicate that rats can differentiate between the sensory consequences of the states produced by CCK, by LiCl, and by ad lib feeding.

Differential effects of intraventricular injections of tachykinin NK1 and NK3 receptor agonists on normal and sham drinking of NaCl by sodium-deficient rats.

The effects of lateral ventricular injections of succinyl-[Asp6, N-Me-Phe8]-substance P (SENK; 25, 100, 200 ng), a tachykinin NK3 receptor agonist, and [Sar9, Met(O2)11]-substance P (Sar Met; 100, 200 ng), an NK1 receptor agonist, on normal (gastric fistula closed) and sham drinking (gastric fistula open) of hypertonic NaCl by sodium-deficient rats were compared. Intraventricular injections of Sar Met had no effect on NaCl intake in either condition. Injections of 100 ng and 200 ng SENK caused an equal suppression of NaCl intake in the 2 fistula conditions. The latency to drink was not affected, but the initial lick rate was significantly lower and decayed more rapidly after 100 ng SENK than after saline or 25 ng SENK. The results show that (a) the tachykinin subtypes are not equally involved in the control of need-induced salt intake; (b) negative feedback from the stomach and distal gastrointestinal tract is not required for intraventricular injections of SENK to suppress sodium appetite; (c) the activation of NK3 receptors decreases the oral excitatory influence of hypertonic NaCl in sodium-deficient rats.

Sodium depletion enhances salt palatability in rats.

Sterotyped fixed action patterns (FAPs) are elicited in rats by oral infusions of taste solutions. These taste-elicited FAPs can be classified as either ingestive or aversive. They reflect the palatability of the taste and can be modified by learning and by the physiological state of the animal. These studies demonstrated that when the physiological state of the rat is altered by sodium depletion, the pattern of FAPs elicited by oral infusions of 0.5 M NaCl shifts from a mixture of ingestive and aversive components (while sodium replete) to exclusively ingestive ones (while sodium deplete). This shift in taste reactivity occurred the first time the rats were made sodium deplete. A similar shift was not observed to accompany infusions of 0.01 M HCl, a taste solution that also elicited mixed ingestive and aversive FAPs. This result suggests that the shift in response to NaCl is not due to a general change in ingestive bias or to a general taste deficit. On the basis of the change in FAPs, it is concluded that the palatability of highly concentrated salt solutions increases in sodium-deplete rats. Such a shift in salt palatability may be instrumental in directing the appetitive behavior of the animal.

The relation of feeding and activity following septal lesions in rats.

The patterns of intake of liquid diet and water were recorded in diet-deprived rats with septal lesions and neurologically intact controls during the first hour of diet access. The occurrences of grooming, resting/sleeping, and exploring were also recorded. Both groups of rats consumed similar amounts of diet in one meal during the 1-hr diet access period. Control rats consumed the meal in one prolonged bout of eating, whereas rats with septal lesions consumed the meal in numerous small bouts of eating. Rats with septal lesions were active for longer periods, exhibiting continuous alternation of brief bouts of eating, drinking, exploring, and resting throughout the meal. In tests in which water was not available during the diet access period, both groups of rats increased their intrameal bout size, but rats with septal lesions still showed much smaller bouts of ingestion than did controls. These data suggest that the small-bout pattern of ingestion may reflect a general disruption in the control of behavioral sequences, rather than processes uniquely related to the regulation of eating or drinking.

Dehydration anorexia in decerebrate rats.

Hypertonic saline (HS) administered intraperitoneally reduced the intake of sucrose solution infused intraorally in tube-fed decerebrate rats, as it did in control animals. Similarly, either intraperitoneal or intravenous HS markedly decreased the intake of laboratory chow by neurologically intact control rats. These observations complement recent findings that lesions of putative osmoreceptors in the ventral diencephalon, which eliminate thirst and blunt pituitary secretion of vasopressin and oxytocin in response to HS in rats, have no apparent effect on the HS-induced inhibition of food intake. Taken together they support previous studies indicating an important role for the caudal brainstem in the central control of food intake and suggest that such brainstem control may also include the inhibition of food intake induced by acute hyperosmolality.

Brain tachykinins and the regulation of salt intake.

The regulation of salt intake is achieved through the coordination of behavioral and physiological responses. Brain neuropeptides, such as the tachykinins, play an important role in orchestrating both of these responses. Intraventricular injections of NK3 receptor agonists, such as senktide, are potent in suppressing salt intake. Experimental results show that intraventricular injections of senktide that suppress salt intake have no effect on the ingestion of other tastes, such as sucrose. The means by which senktide suppresses salt intake was investigated in a series of experiments. Taste reactivity and lick rate analyses suggest that the activation of NK3 receptors reduces salt intake by modulating the oral-stimulating property of salt taste.